Mycoplasma pneumoniae is an important cause of human respiratory disease and has been implicated in many acute and chronic airway pathologies including asthma, as well as in a range of extra pulmonary manifestations. The diagnosis of M. pneumoniae has been difficult for many reasons, including their highly fastidious nature. Our discovery of the ADP-ribosylating and vacuolating CARDS TX has enabled us to develop molecular and immunological assays (in collaboration with the diagnostic company Alere) to detect M. pneumoniae based upon the CARDS TX target. We have published these initial assays that continue to evolve in terms of reagent and format optimization. Our long-term objectives of the Diagnostic Core Laboratory will be to provide novel diagnostic assays that are highly sensitive and specific for rapid detection of Mycoplasma pneumoniae, based upon CARDS TX as target. This will allow us to monitor and evaluate the frequency and impact of M. pneumoniae and CARDS TX on asthma-related symptomatology in pediatric and adult patients and pulmonary inflammation, asthma development and pathogenesis in the infected murine model. Our goal is to evaluate specimens such as nasal swabs, throat swabs, sputum and sera from acute and chronic pediatric and adult asthma and control patients to determine the incidence of M. pneumoniae and to correlate the presence of this organism, and CARDS TX-specific antibodies, to pulmonary outcomes using longitudinal assessments. Our hypothesis is that M. pneumoniae through its bona fide virulence determinant, CARDS TX, elicits and exacerbates allergic airway Inflammation in pediatric and adult populations. By accumulating meaningful data from both initial and longitudinal studies of patients and healthy controls, and from experimental data obtained through the murine model of infection and intoxication, we will generate improved diagnostic information which will lead to effective and enhanced treatment modalities. Our specific aims are as follows: (1) collection and transport of clinical and experimental samples in conjunction with Patient Core and Project 1;(2) process clinical and experimental samples for diagnostic analysis;(3) perform CARDS TX ELISA on all serum samples, both clinical and experimental;(4) evaluate biological samples for levels of CARDS TX protein using a unique antigen capture assay;and (5) perform CARDS TX quantitative real time PCR on samples as described.